The carboxy-terminal αN helix of the archaeal XerA tyrosine recombinase is a molecular switch to control site-specific recombination

PLoS One. 2013 May 7;8(5):e63010. doi: 10.1371/journal.pone.0063010. Print 2013.

Abstract

Tyrosine recombinases are conserved in the three kingdoms of life. Here we present the first crystal structure of a full-length archaeal tyrosine recombinase, XerA from Pyrococcus abyssi, at 3.0 Å resolution. In the absence of DNA substrate XerA crystallizes as a dimer where each monomer displays a tertiary structure similar to that of DNA-bound Tyr-recombinases. Active sites are assembled in the absence of dif except for the catalytic Tyr, which is extruded and located equidistant from each active site within the dimer. Using XerA active site mutants we demonstrate that XerA follows the classical cis-cleavage reaction, suggesting rearrangements of the C-terminal domain upon DNA binding. Surprisingly, XerA C-terminal αN helices dock in cis in a groove that, in bacterial tyrosine recombinases, accommodates in trans αN helices of neighbour monomers in the Holliday junction intermediates. Deletion of the XerA C-terminal αN helix does not impair cleavage of suicide substrates but prevents recombination catalysis. We propose that the enzymatic cycle of XerA involves the switch of the αN helix from cis to trans packing, leading to (i) repositioning of the catalytic Tyr in the active site in cis and (ii) dimer stabilisation via αN contacts in trans between monomers.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Apoenzymes / chemistry
  • Apoenzymes / metabolism
  • Base Sequence
  • Crystallography, X-Ray
  • DNA, Archaeal / genetics*
  • Models, Molecular
  • Protein Multimerization
  • Protein Structure, Quaternary
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Pyrococcus abyssi / enzymology*
  • Pyrococcus abyssi / genetics
  • Recombinases / chemistry*
  • Recombinases / metabolism*
  • Recombination, Genetic*
  • Tyrosine*

Substances

  • Apoenzymes
  • DNA, Archaeal
  • Recombinases
  • Tyrosine

Grants and funding

This work was supported by funds from the Centre National de la Recherche Scientifique and the University Paris-Sud (UMR8619 and UMR8621). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.